Synthesis of Tumor-Associated Le(a)Le(x) Hexasaccharides: Instability of a Thiol-Containing Oligosaccharide in Mass Spectrometry and Hypermetalation Detected by ESI FAIMS.

We report the efficient synthesis of three analogues of the tumor-associated carbohydrate antigen Le(a)Le(x). This hexasaccharide was prepared as a soluble inhibitor hexyl glycoside, as a 6-aminohexyl glycoside for conjugation to proteins, and as a 6-thiohexyl glycoside for immobilization to a gold surface. These three analogues were obtained from a common hexasaccharide intermediate and isolated pure following efficient deprotection reactions that involved metal-dissolving conditions. While all other intermediates and analogues gave the expected molecular ions in ESI HRMS, the 6-thiohexyl glycoside final compound gave a complex spectrum in which no signal matched the molecular ion. Using ESI FAIMS HRMS, we were able to prevent ion dissociation reactions and obtained high quality spectral data. The ions detected could be characterized unambiguously from their accurate masses and gave insight into the behavior of the thiohexyl analogue in the gas phase. These results indicate that the 6-thiohexyl glycoside lost water and led to the formation of "hypermetalated" species which we propose are cyclic.